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1.
J Clin Immunol ; 39(1): 81-89, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30607663

RESUMO

The association of immunodeficiency-related vaccine-derived rubella virus (iVDRV) with cutaneous and visceral granulomatous disease has been reported in patients with primary immunodeficiency disorders (PIDs). The majority of these PID patients with rubella-positive granulomas had DNA repair disorders. To support this line of inquiry, we provide additional descriptive data on seven previously reported patients with Nijmegen breakage syndrome (NBS) (n = 3) and ataxia telangiectasia (AT) (n = 4) as well as eight previously unreported patients with iVDRV-induced cutaneous granulomas and DNA repair disorders including NBS (n = 1), AT (n = 5), DNA ligase 4 deficiency (n = 1), and Artemis deficiency (n = 1). We also provide descriptive data on several previously unreported PID patients with iVDRV-induced cutaneous granulomas including cartilage hair hypoplasia (n = 1), warts, hypogammaglobulinemia, immunodeficiency, myelokathexis (WHIM) syndrome (n = 1), MHC class II deficiency (n = 1), Coronin-1A deficiency (n = 1), X-linked severe combined immunodeficiency (X-SCID) (n = 1), and combined immunodeficiency without a molecular diagnosis (n = 1). At the time of this report, the median age of the patients with skin granulomas and DNA repair disorders was 9 years (range 3-18). Cutaneous granulomas have been documented in all, while visceral granulomas were observed in six cases (40%). All patients had received rubella virus vaccine. The median duration of time elapsed from vaccination to the development of cutaneous granulomas was 48 months (range 2-152). Hematopoietic cell transplantation was reported to result in scarring resolution of cutaneous granulomas in two patients with NBS, one patient with AT, one patient with Artemis deficiency, one patient with DNA Ligase 4 deficiency, one patient with MHC class II deficiency, and one patient with combined immunodeficiency without a known molecular etiology. Of the previously reported and unreported cases, the majority share the diagnosis of a DNA repair disorder. Analysis of additional patients with this complication may clarify determinants of rubella pathogenesis, identify specific immune defects resulting in chronic infection, and may lead to defect-specific therapies.


Assuntos
Reparo do DNA/genética , Granuloma/complicações , Granuloma/virologia , Síndromes de Imunodeficiência/complicações , Vírus da Rubéola/patogenicidade , Dermatopatias/etiologia , Dermatopatias/virologia , Adolescente , Ataxia Telangiectasia/genética , Ataxia Telangiectasia/virologia , Criança , Pré-Escolar , Feminino , Granuloma/genética , Cabelo/anormalidades , Cabelo/virologia , Transplante de Células-Tronco Hematopoéticas/métodos , Doença de Hirschsprung/genética , Doença de Hirschsprung/virologia , Humanos , Síndromes de Imunodeficiência/genética , Síndromes de Imunodeficiência/virologia , Masculino , Síndrome de Quebra de Nijmegen/genética , Síndrome de Quebra de Nijmegen/virologia , Osteocondrodisplasias/congênito , Osteocondrodisplasias/genética , Osteocondrodisplasias/virologia , Rubéola (Sarampo Alemão)/genética , Rubéola (Sarampo Alemão)/virologia , Pele/virologia , Dermatopatias/genética , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/genética , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/virologia
2.
Orphanet J Rare Dis ; 13(1): 207, 2018 11 16.
Artigo em Inglês | MEDLINE | ID: mdl-30445974

RESUMO

BACKGROUND: Patients with cartilage-hair hypoplasia (CHH), a rare metaphyseal chondrodysplasia, manifest severe growth failure, variable immunodeficiency and increased risk of malignancies. The impact of CHH on gynecologic and reproductive health is unknown. Vulnerability to genital infections may predispose CHH patients to prolonged human papillomavirus (HPV) infections potentially leading to cervical, vaginal and vulvar cancer. METHODS: We carried out gynecologic evaluation, pelvic ultrasound and laboratory assessment in 19 women with genetically confirmed CHH. All patients were clinically examined and retrospective data were collected from hospital records. RESULTS: The women ranged in age from 19.2 to 70.8 years (median 40.8 years) and in height from 103 to 150 cm (median 123 cm). All women had undergone normal pubertal development as assessed by breast development according to Tanner scale and by age of menarche (mean 12.5 yrs., range 11-14 yrs). Despite significant short stature and potentially small pelvic diameters, a well-developed uterus with fairly normal size and shape was found by pelvic ultrasound in most of the patients. Ovarian follicle reserve, assessed by ultrasound was normal in relation to age in all premenopausal women it could be assessed (12 cases). Anti-Müllerian hormone was normal in relation to age in 17 women (89%). HPV was detected in 44% (8/18) and three women carried more than one HPV serotype; findings did not associate with immunological parameters. Three patients had a concurrent cell atypia in Pap smear. CONCLUSIONS: Pubertal development, reproductive hormones and ovarian structure and function were usually normal in women with CHH suggesting fairly normal reproductive health. However, the immunodeficiency characteristic to CHH may predispose the patients to HPV infections. High prevalence of HPV infections detected in this series highlights the importance of careful gynecologic follow up of these patients.


Assuntos
Cabelo/anormalidades , Doença de Hirschsprung/patologia , Doença de Hirschsprung/virologia , Síndromes de Imunodeficiência/patologia , Síndromes de Imunodeficiência/virologia , Osteocondrodisplasias/congênito , Papillomaviridae/patogenicidade , Adulto , Idoso , Feminino , Genótipo , Cabelo/patologia , Cabelo/virologia , Doença de Hirschsprung/genética , Humanos , Síndromes de Imunodeficiência/genética , Pessoa de Meia-Idade , Osteocondrodisplasias/genética , Osteocondrodisplasias/patologia , Osteocondrodisplasias/virologia , Folículo Ovariano/metabolismo , Folículo Ovariano/patologia , Estudos Retrospectivos , Sorogrupo
3.
J Infect Dis ; 211(9): 1437-46, 2015 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-25387582

RESUMO

BACKGROUND: Several new polyomaviruses have been discovered in the last decade, including Merkel cell polyomavirus (MCPyV). Little is known about the natural history of the more recently discovered polyomaviruses. We estimated the incidence, prevalence, and persistence of 9 polyomaviruses (MCPyV, BK polyomavirus, KI polyomavirus, JC polyomavirus, WU polyomavirus, Human polyomavirus 6 [HPyV6], HPyV7, HPyV9, and Trichodysplasia spinulosa-associated polyomavirus) and examined factors associated with MCPyV infection in a prospective cohort of 209 men initially enrolled in the HPV Infection in Men (HIM) study. METHODS: Participants enrolled at the US site of the HIM study were recruited into a substudy of cutaneous viral infections and followed for a median of 12.6 months. Eyebrow hair and normal skin swab specimens were obtained at each study visit, and the viral DNA load was measured using multiplex polymerase chain reaction. RESULTS: MCPyV infection showed the highest prevalence (65.1% of normal skin swab specimens and 30.6% of eyebrow hair specimens), incidence (81.7 cases per 1000 person-months among normal skin swab specimens, and 24.1 cases per 1000 person-months among eyebrow hair specimens), and persistence (85.8% of normal skin swab specimens and 58.9% of eyebrow hair specimens) among all polyomaviruses examined. Age of >44 years (odds ratio [OR], 2.11; 95% confidence interval [CI], 1.03-4.33) and Hispanic race (OR, 2.64; 95% CI, 1.01-6.88) were associated with an increased prevalence of MCPyV infection in eyebrow hair and normal skin swab specimens, respectively. CONCLUSION: MCPyV infection is highly prevalent in adults, with age and race being predisposing factors.


Assuntos
Infecções por Polyomavirus/virologia , Polyomavirus/classificação , Adolescente , Adulto , Idoso , Cabelo/virologia , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Infecções por Polyomavirus/epidemiologia , Prevalência , Fatores Sexuais , Pele/virologia , Estados Unidos/epidemiologia , Adulto Jovem
4.
J Am Acad Dermatol ; 71(1): 108-15.e1, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24612648

RESUMO

BACKGROUND: Correlating human papillomavirus (HPV) type with the clinical and histopathological features of skin lesions (from genital and nongenital sites) can present a diagnostic challenge. OBJECTIVE: In this study, HPV infection patterns were correlated with pathology and clinical presentation in lesional and nonlesional body sites from a young patient with a primary T-cell immunodeficiency. METHODS: HPV infection was evaluated at both DNA and protein levels by polymerase chain reaction and immunohistochemistry. RESULTS: The patient's genital lesions were caused exclusively by α-genotypes (high-risk type HPV-51 in the anal and low-risk type HPV-72 in the penile condylomas). The opposite was true for the skin lesions, which were infected by ß-genotypes alone (HPV-8 and HPV-24). HPV-24 was the predominant type in terms of viral load, and the only one found in productive areas of infection. The patient had already developed high-grade dysplasia in the anal condyloma-like lesions, and showed areas of early-stage dysplasia in the lesions caused by the ß-genotype HPV-24. LIMITATIONS: The basic origin of the immunodeficiency is not yet defined. CONCLUSION: These findings provide proof of principle that both α- and ß-genotypes can cause overt dysplastic lesions when immunosurveillance is lost, which is not restricted to epidermodysplasia verruciformis.


Assuntos
Condiloma Acuminado/virologia , Síndromes de Imunodeficiência/complicações , Infecções por Papillomavirus/patologia , Adulto , Alphapapillomavirus/classificação , Alphapapillomavirus/genética , Betapapillomavirus/genética , Betapapillomavirus/isolamento & purificação , DNA Viral/análise , Citometria de Fluxo , Genótipo , Cabelo/virologia , Humanos , Síndromes de Imunodeficiência/virologia , Linfopenia/imunologia , Masculino , Membrana Mucosa/virologia , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/virologia , Reação em Cadeia da Polimerase em Tempo Real
5.
J Vet Sci ; 13(3): 323-5, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23000590
6.
J Gen Virol ; 93(Pt 8): 1774-9, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22552941

RESUMO

Recent studies indicate that human papillomaviruses (HPVs) from the genera Betapapillomavirus and Gammapapillomavirus are abundant in the human oral cavity. We report the cloning and characterization of a 7304 bp HPV120 genome from the oral cavity that is related most closely to HPV23 (L1 ORF, 83.7 % similarity), clustering it in the genus Betapapillomavirus (ß-PV). HPV120 contains five early and two late genes, but no E5 ORF. HPV120 was detected from heterogeneous human biological niches, including the oral cavity, eyebrow hairs, anal canal and penile, vulvar and perianal warts. Characterization of the clinical spectrum of HPV120 infections indicates a broader spectrum of epithelial tropism than appreciated previously for HPV types from the genus ß-PV.


Assuntos
Betapapillomavirus/classificação , Betapapillomavirus/genética , Tropismo Viral/fisiologia , Canal Anal/virologia , Condiloma Acuminado/virologia , Regulação Viral da Expressão Gênica , Genoma Viral , Cabelo/virologia , Humanos , Dados de Sequência Molecular , Boca/virologia , Infecções por Papillomavirus/virologia , Filogenia , Polimorfismo de Nucleotídeo Único
7.
J Vet Sci ; 12(3): 295-7, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21897105

RESUMO

The purpose of this study was to determine whether manually plucked hairs might serve as an alternative sample for a quantitative real time polymerase chain reaction (qRT-PCR) testing. Twenty three, 1~3 week old, non-bovine viral diarrhea virus (BVDV) vaccinated calves, found to be positive for BVDV by immunohistochemical staining, were selected and hairs were manually plucked from the ear. qRT-PCR was performed on samples consisting of more than 30 hairs (30~100) and whole blood. All 23 animals were positive for the virus by qRT-PCR performed on the whole blood and when samples of more than 30 hairs were assayed. Additionally, qRT-PCR was performed on groups of 10 and 20 hairs harvested from 7 out of 23 immunohistochemical staining-positive calves. When groups of 20 and 10 hairs were tested, 6 and 4 animals, respectively, were positive for the virus.


Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/diagnóstico , Vírus da Diarreia Viral Bovina Tipo 1/isolamento & purificação , Vírus da Diarreia Viral Bovina Tipo 2/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Animais , Anticorpos Antivirais/análise , Doença das Mucosas por Vírus da Diarreia Viral Bovina/sangue , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Bovinos , Vírus da Diarreia Viral Bovina Tipo 1/genética , Vírus da Diarreia Viral Bovina Tipo 2/genética , Cabelo/virologia , RNA Viral/química , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos
8.
J Vet Diagn Invest ; 22(3): 451-4, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20453226

RESUMO

A wildlife sanctuary presented an adult female cottontail rabbit (Sylvilagus spp.), age unknown, to the Colorado State University Pathology service for postmortem examination. Gross examination revealed numerous pigmented wartlike lesions arising from the skin of the head surrounding the ears, eyes, nares, mouth, and dorsum. Masses were firm, friable, and easily detached from the underlying skin. Differential diagnoses included Cottontail rabbit papillomavirus, Rabbit fibroma virus, and Myxoma virus. Histological examination revealed multiple papillary masses lined by stratified squamous epithelial cells with central cores of fibrovascular connective tissue and parakeratotic hyperkeratosis. Cells of the Stratum spinosum were frequently swollen with abundant perinuclear, cytoplasmic, clearing, and occasional intranuclear basophilic, glassy, spherical inclusions up to 3 microm in diameter. The lesions were consistent with Cottontail rabbit papillomavirus infection. Papilloma virus antigens were identified by immunohistochemistry. In addition, papillomavirus particles were identified by transmission electron microscopy within Langerhans cells of the epidermis, suggesting a unique mechanism for systemic dissemination of the virus. The present case report highlights the finding of viral particles within the Langerhans cells and suggests a novel mechanism of pathogenesis.


Assuntos
Papillomavirus de Coelho Cottontail/isolamento & purificação , Células de Langerhans/virologia , Infecções por Papillomavirus/veterinária , Animais , Animais Selvagens/virologia , Feminino , Cabelo/patologia , Cabelo/virologia , Imuno-Histoquímica , Células de Langerhans/patologia , Infecções por Papillomavirus/patologia , Coelhos , Pele/patologia , Pele/virologia
9.
Int J Cancer ; 126(11): 2614-21, 2010 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-19856311

RESUMO

Human papillomaviruses from the genus beta (betaPV) are a possible cause of cutaneous squamous cell carcinoma (SCC). We compared the betaPV infections in SCC and in sets of cutaneous tissues collected from a series of individual SCC patients to determine concordance and to assess the adequacy of eyebrow hairs as noninvasive markers of betaPV infection. Biopsies of SCC tumors, perilesional tissue, normal skin from the mirror image of nonfacial SCC and plucked eyebrow hairs were collected from 21 patients with incident SCC living in Queensland, Australia. These were tested for the presence of DNA from 25 different betaPV types. Overall prevalence of betaPV was high in every sample type, ranging from 81% to 95%. The median number of types was significantly higher in the SCC tumour (6), perilesional skin (5) and eyebrow hairs (5) than in normal skin (2). Comparing SCC tissue with other sample types within patients showed 63 overlapping infections with eyebrow hairs (71%; 95% CI: 60-80); 56 with perilesional skin samples (63%; 95% CI: 52-73) and 23 with normal skin samples (26%; 95% CI: 17-36). The sensitivity of eyebrow hair testing for detection of betaPV in the tumor was 82% (95% CI: 57-96) with concordance defined as 50% of betaPV types in common and 29% (95% CI: 10-56) for 100% concordance. These findings support the concept that perilesional skin represents an area of field change involving betaPV preceding SCC development and indicate that eyebrow hairs can serve to some degree as an easily collected marker of tumor betaPV status in epidemiological studies.


Assuntos
Betapapillomavirus/isolamento & purificação , Carcinoma de Células Escamosas/virologia , Sobrancelhas/virologia , Infecções por Papillomavirus/epidemiologia , Neoplasias Cutâneas/virologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Austrália , DNA/genética , DNA/isolamento & purificação , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Genótipo , Cabelo/virologia , Humanos , Masculino , Pessoa de Meia-Idade , Infecções por Papillomavirus/genética , Reação em Cadeia da Polimerase , Pele/virologia
10.
J Infect Dis ; 199(9): 1270-4, 2009 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-19301978

RESUMO

Human papillomaviruses (HPVs) were detected in 69 (43.7%) of 158 and in 7 (4.5%) of 155 anogenital hairs obtained from 53 patients with genital warts (GWs) and from 53 age-matched healthy control subjects, respectively. At least 1 hair sample was positive for 69.8% of patients and for 13.2% of control subjects. For patients, HPV was detected in 64.2%, 39.6%, and 26.9% of hairs plucked from the pubic, scrotal, and perianal regions, respectively. For 91.9% of patients, the same HPV genotype was identified in GWs and hairs from at least 1 sampling site. Having GWs was found to be strongly associated with the presence in anogenital hairs of the HPV genotype causing the GWs (range of odds ratios, 13.0-20.0).


Assuntos
Alphapapillomavirus/isolamento & purificação , Condiloma Acuminado/virologia , Cabelo/virologia , Adolescente , Adulto , Alphapapillomavirus/crescimento & desenvolvimento , Canal Anal/virologia , Condiloma Acuminado/psicologia , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Prepúcio do Pênis/virologia , Humanos , Masculino , Pessoa de Meia-Idade , Pênis/virologia , Reação em Cadeia da Polimerase , Escroto/virologia , Pele/virologia , Vulva/virologia , Adulto Jovem
11.
J Gen Virol ; 88(Pt 10): 2670-8, 2007 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17872518

RESUMO

The high incidence of multiple wart formation and skin cancer in organ-transplant recipients, as well as the question of an involvement of papillomaviruses in a variety of human cancers, require a model system for papillomavirus infections in immunocompetent animals. Such an in vivo model is represented by the multimammate rat Mastomys coucha, which is infected with Mastomys natalensis papillomavirus (MnPV). MnPV primarily induces benign skin tumours, such as papillomas and keratoacanthomas. Here, the incidence of MnPV infections in different skin areas and various organs is described. In situ hybridization showed that hair follicle cells were positive for viral DNA and that the amount of MnPV in normal skin may be considered a predictor for the development of skin tumours. MnPV infection is not restricted to the skin, but can also be detected in inner organs. As the blood and the lymphatic system were temporarily also found to be virus-positive, a haematogenic propagation of MnPV can be assumed. However, MnPV is apparently not transmitted through the germ line, as fetuses and newborns lack viral DNA, despite infection of their mothers. In conclusion, M. coucha is not only useful to study papillomavirus-induced skin carcinogenesis, but may also serve as a model to identify additional, still unknown target cells of papillomavirus infections and the potential pathological impact.


Assuntos
Murinae/virologia , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/patologia , Neoplasias Cutâneas/virologia , Animais , Sangue/virologia , Southern Blotting , DNA Viral/isolamento & purificação , Embrião de Mamíferos/virologia , Cabelo/virologia , Neoplasias de Cabeça e Pescoço/virologia , Papillomavirus Humano 16/patogenicidade , Papillomavirus Humano 18/patogenicidade , Humanos , Hibridização In Situ , Papillomaviridae/genética , Reação em Cadeia da Polimerase , RNA Viral/genética , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa
12.
J Gen Virol ; 88(Pt 5): 1489-95, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-17412978

RESUMO

Infections with human papillomaviruses (HPVs) belonging to the genus Betapapillomavirus have been linked to the development of non-melanoma skin cancer. Although persistence is expected, systematic investigation of this aspect of betapapillomavirus (beta-PV) infection has not been conducted. This study investigated the prevalence and persistence of 25 known beta-PV types in the skin of immunocompetent individuals. Over a 2 year period, eight consecutive plucked eyebrow hair samples taken from 23 healthy individuals were analysed for the presence of beta-PV DNA. Using a recently published general beta-PV PCR and genotyping method, 61% of the individuals were beta-PV DNA positive for one or more types at intake, whereas during follow-up this percentage rose to 96%. HPV23 was the most frequently detected beta-PV type. Type-specific beta-PV DNA was detected over 6 months or longer in 74% of the individuals. In 57% of the individuals, DNA from multiple beta-PV types was detected simultaneously for 6 months or longer. When the detection intervals of all beta-PV type-specific infections in the study population were considered, a substantial proportion, 48%, lasted at least half a year. The consistent beta-PV patterns found over time in most individuals strongly suggested that beta-PV DNA detection in plucked eyebrow hairs reveals true beta-PV infection. If the minimum interval of detection was set at 6 months, persistent beta-PV infections were found in the majority of the study population (74%).


Assuntos
Betapapillomavirus/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , Pele/virologia , Adulto , Betapapillomavirus/genética , Cabelo/virologia , Humanos , Imunocompetência , Hibridização de Ácido Nucleico , Reação em Cadeia da Polimerase , Valores de Referência
14.
Transplant Proc ; 38(9): 3066-9, 2006 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17112901

RESUMO

Xenotransplantation of porcine organs has the potential to overcome the current critical shortage of allogenic organs for transplantation in humans. However, the existence of porcine endogenous retroviruses (PERVs) presents a problem for the clinical use of xenografts from pigs. In an attempt to understand the molecular characteristics of PERVs, we cloned the PERV env gene from six pig breeds (ie, Berkshire, Duroc, Landrace, Yorkshire, and two types of miniature pigs) in Korea. A total of 141 env clones were isolated and their sequences were analyzed. Phylogenetic analyses of these genes revealed the presence of PERVs, from both classes A and B, in 54% and 46% of the env clones, respectively. Among these clones, 37 isolates had the correct open reading frame (ORF; 27 clones in subclass A and 10 clones in subclass B), while the others had premature termination. These PERV nucleotide sequences can be used in a database for comparisons of PERV distribution among different pig breeds and for monitoring PERV infection using isolates with functional ORFs. Recombinant envelope of subclass A and B with functional ORF was expressed by vaccinia virus systems. Additionally isolated env clones can be used for various experiments, such as PERV control and infectivity tests, and may enhance the understanding of molecular mechanisms through pseudotyped PERV viruses.


Assuntos
Retrovirus Endógenos/genética , Cabelo/virologia , Suínos/virologia , Proteínas do Envelope Viral/genética , Animais , Retrovirus Endógenos/isolamento & purificação , Coreia (Geográfico) , Porco Miniatura/virologia
15.
J Med Virol ; 78(12): 1673-8, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17063512

RESUMO

A total of 150 specimens of anogenital hairs plucked from the scrotal, pubic, and perianal region of 51 immunocompetent healthy male individuals were tested for the presence of beta-papillomaviruses (beta-HPV) using the nested M(a)/H(a) polymerase chain reaction. Beta-HPV were found in a total of 38 (25.3%) of 150 hair samples. According to the sampling sites, beta-HPV were detected in 18/51 (35.3%), 13/50 (26.0%), and 7/49 (14.3%) plucked hair samples obtained from the pubic, scrotal, and perianal region, respectively. The prevalence of beta-HPV in the plucked pubic hairs was significantly higher than in the perianal hairs (P = 0.013). In contrast, the difference in the prevalence of beta-HPV in the pubic and scrotal hairs as well as in scrotal and perianal hairs did not reach statistical significance (P = 0.302 and P = 0.227, respectively). The difference in the lifetime-cumulative sun exposure is the most likely explanation for the differences obtained on beta-HPV prevalence. Beta-HPV genotype HPV-38 was detected most frequently, followed by HPV-36, HPV-15, and HPV-14D. In addition to the beta-HPV recognized officially five partial DNA sequences suggesting putative new HPV genotypes were identified.


Assuntos
Canal Anal/virologia , Betapapillomavirus/classificação , Betapapillomavirus/isolamento & purificação , Cabelo/virologia , Imunocompetência , Infecções por Papillomavirus/epidemiologia , Escroto/virologia , Adolescente , Adulto , Betapapillomavirus/genética , DNA Viral/análise , DNA Viral/isolamento & purificação , Genótipo , Humanos , Masculino , Dados de Sequência Molecular , Infecções por Papillomavirus/virologia , Filogenia , Prevalência , Sínfise Pubiana , Análise de Sequência de DNA
16.
J Clin Microbiol ; 44(5): 1792-800, 2006 May.
Artigo em Inglês | MEDLINE | ID: mdl-16672409

RESUMO

Human papillomavirus can be detected by amplification of viral DNA. A novel one-step PCR (PM-PCR) was evaluated for amplification of a 117-bp fragment from the E1 region. It permitted ultrasensitive detection of all 25 known human papillomavirus genotypes from the beta-papillomavirus genus. The intra- and intertypic sequence variations of the 77-bp interprimer region were studied. Genotype-specific probes as well as general probes were selected for the 25 established beta-papillomavirus types, and a reverse hybridization assay (RHA) was developed (PM-PCR RHA method). The analytical sensitivity of the PM-PCR RHA method was 10 to 100 viral genomes. The one-step PM-PCR turned out to be more sensitive than the previously described nested MaHa-PCR for beta-papillomavirus detection. The PM-PCR RHA method was able to detect and identify beta-papillomavirus types in frozen patient material as well as in poorly amplifiable material such as formalin-fixed, paraffin-embedded skin biopsy specimens. Inter- and intralaboratory variability experiments showed that the reproducibility of the assay was very high. In conclusion, the one-step PM-PCR together with the RHA allows extremely sensitive, specific, and reproducible detection of beta-papillomavirus DNA as well as reliable identification of beta-papillomavirus genotypes in both fresh and paraffin-embedded patient material.


Assuntos
DNA Viral/genética , DNA Viral/isolamento & purificação , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Sequência de Bases , Estudos de Casos e Controles , Genótipo , Cabelo/virologia , Humanos , Laboratórios , Dados de Sequência Molecular , Papillomaviridae/classificação , Inclusão em Parafina , Reação em Cadeia da Polimerase/estatística & dados numéricos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Homologia de Sequência do Ácido Nucleico , Virologia/métodos , Virologia/estatística & dados numéricos
17.
Methods Mol Med ; 119: 115-27, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-16350401

RESUMO

Epidemiological studies, which address the role of human papillomavirus (HPV) in the pathogenesis of (pre)malignant cutaneous lesions, focus on the HPV B1 subgroup comprising the so-called epidermodysplasia verruciformis (EV)-associated HPV types. To detect and type HPV DNA in human materials, Polymerase Chain Reaction (PCR)-based assays are used. In this chapter, a nested, broad-spectrum PCR method using a mixture of primers and a type-specific PCR using specific primers are described. The broad-spectrum PCR detects the B1 subgroup of HPV types. HPV typing is performed by sequence analysis of the PCR product. The type-specific PCR detects and types HPV 5a, 8, 15, 17, 20, 24, 36, and 38. These HPV types are representative of the B1 subgroup, because they are evenly distributed over the phylogenetic tree of the B1 subgroup.


Assuntos
DNA Viral/análise , Papillomaviridae/classificação , Papillomaviridae/isolamento & purificação , Lesões Pré-Cancerosas/virologia , Dermatopatias Virais/diagnóstico , Sequência de Bases , Biópsia , Clonagem Molecular/métodos , Primers do DNA , DNA Viral/genética , Cabelo/citologia , Cabelo/virologia , Humanos , Papillomaviridae/genética , Reação em Cadeia da Polimerase/métodos , Lesões Pré-Cancerosas/diagnóstico , Pele/citologia , Pele/virologia
18.
J Clin Microbiol ; 43(11): 5581-7, 2005 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16272490

RESUMO

The beta and gamma genera of papillomaviruses consist of epidermodysplasia verruciformis-related human papillomaviruses (HPVs) and phylogenetically related cutaneous HPVs. Here, we have developed a consensus primer PCR assay and reverse line blot typing system coupled thereto (referred to as beta and gamma cutaneous HPV PCR [BGC-PCR]) for detection and typing of 24 beta and gamma HPVs (HPV types 4, 5, 8, 9, 12, 14, 15, 17, 19, 20, 21, 22, 23, 24, 25, 36, 37, 38, 47, 48, 49, 50, 60, and 65). Because the HPV-specific PCR products are only 72 bp in size, the system is suitable for formalin-fixed, paraffin-embedded specimens and other samples in which the DNA is of suboptimal quality. This system was able to detect and type as little as 100 ag to 1 fg HPV DNA per reaction (depending on the HPV type) in a background of 100 ng human DNA without any cross-reactivity between the tested types. Beta and gamma HPVs were detected in DNA extracted from plucked eyebrow hairs of 31 of 34 renal transplant recipients. In addition, formalin-fixed, paraffin-embedded specimens from nonmelanoma skin tumors of renal transplant recipients (n = 25) and immunocompetent individuals (n = 15) scored BGC-PCR positive in 21 and 6 cases, respectively, with HPV type 5 (HPV5) and HPV8 being the predominant types. The data indicate that this method can be a valuable, user-friendly tool for the detection and typing of cutaneous HPV in clinical specimens and may have implications for future monitoring of vaccines or alternative treatment modalities for diseases caused by these cutaneous HPVs.


Assuntos
Papillomaviridae/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Verrugas/diagnóstico , Sequência de Bases , Biópsia , Primers do DNA/genética , Cabelo/virologia , Humanos , Imunocompetência , Transplante de Rim , Dados de Sequência Molecular , Papillomaviridae/genética , Sensibilidade e Especificidade , Alinhamento de Sequência , Pele/patologia , Pele/virologia , Verrugas/etiologia , Verrugas/patologia , Verrugas/virologia
19.
J Med Virol ; 73(3): 465-73, 2004 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-15170644

RESUMO

We obtained 7,566 peripheral blood mononuclear cell (PBMC) samples from 2,332 individuals and screened them for human herpesvirus infection. We identified five individuals who persistently harbored high copy numbers of human herpesvirus 6 (HHV-6) DNA in their PBMCs. HHV-6 DNA was also detected in other somatic tissues of these individuals. Five additional cases were identified among their family members. For two of these families, chromosomally integrated HHV-6 DNA (CIHHV-6) was detected in the PBMCs by fluorescence in situ hybridization. The prevalence of CIHHV-6 among all the subjects was 0.21%. The HHV-6 DNA was variant B in four families and variant A in one family. Antibodies to immediate early antigen and glycoprotein B were detected in 57 and 14% of individuals with CIHHV-6 and in 0 and 60% of healthy volunteers without CIHHV-6, respectively. HHV-6 could not be isolated from PBMCs with CIHHV-6. These cases shared no clinical features, and included three healthy individuals. Our data suggest that CIHHV-6 is rare but detectable in the general population and that hereditary transmission is one of the routes of HHV-6 transmission.


Assuntos
DNA Viral/análise , Herpesvirus Humano 6/genética , Transmissão Vertical de Doença Infecciosa , Leucócitos Mononucleares/virologia , Infecções por Roseolovirus/transmissão , Infecções por Roseolovirus/virologia , Integração Viral , Anticorpos Antivirais/sangue , Cromossomos Humanos/virologia , Feminino , Cabelo/virologia , Herpesvirus Humano 6/isolamento & purificação , Humanos , Proteínas Imediatamente Precoces/imunologia , Hibridização in Situ Fluorescente , Masculino , Mucosa Bucal/virologia , Linhagem , Faringe/virologia , Fosfoproteínas/imunologia , Reação em Cadeia da Polimerase , Proteínas do Envelope Viral/imunologia
20.
Arch Dermatol ; 140(3): 317-24, 2004 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15023775

RESUMO

BACKGROUND: Patients with psoriasis treated with psoralen-UV-A (PUVA) are at increased risk of skin cancer; however, the exact causes of this increased incidence are not well understood. It has been suggested that PUVA may increase expression of the tumorigenic agent human papillomavirus (HPV) in skin by directly stimulating virus replication, immune suppression, or both, thereby leading to skin cancer formation. OBJECTIVE: To determine whether HPV DNA prevalence in the skin is increased after long-term PUVA treatment. DESIGN: Screening for the presence of HPV sequences in DNA isolated from plucked body hairs of patients with psoriasis with a history of PUVA exposure and a history of skin cancer (group A), PUVA exposure and no history of skin cancer (group B), and no PUVA exposure and no history of skin cancer (group C). SETTING: University hospital. PATIENTS AND METHODS: Hair samples were obtained from 81 patients with psoriasis (56 men and 25 women; mean age, 52 years), including 16 in group A (mean number of PUVA exposures, 702), 35 in group B (mean number of PUVA exposures, 282), and 30 in group C. DNA was isolated from the hair samples and analyzed by polymerase chain reaction with the use of 2 nested primer systems specific for epidermodysplasia verruciformis-associated or related and genital or mucosal virus types, respectively. RESULTS: The rate of HPV DNA positivity was significantly higher in groups A (73% [11/15]) and B (69% [24/35]) than in group C (36% [10/28]) (A + B vs C, P =.009; chi(2) test; age adjusted). Conclusion The prevalence of HPV in the skin (hair follicles) is increased in patients with psoriasis who have a history of PUVA exposure.


Assuntos
Cabelo/virologia , Terapia PUVA/efeitos adversos , Papillomaviridae/isolamento & purificação , Psoríase/epidemiologia , Psoríase/virologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Áustria/epidemiologia , Estudos de Casos e Controles , DNA Viral/análise , Feminino , Ficusina/administração & dosagem , Humanos , Masculino , Pessoa de Meia-Idade , Papillomaviridae/classificação , Papillomaviridae/genética , Fármacos Fotossensibilizantes/administração & dosagem , Prevalência , Psoríase/tratamento farmacológico
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